Off-note blocking sensory organic compounds

ABSTRACT

Disclosed are compounds that block off-notes in consumables and methods of blocking off-notes in consumables including off-notes provided by artificial sweeteners including aspartame, saccharin, acesulfame K (Acesulfame potassium), sucralose and cyclamate; and including stevioside, swingle extract, glyccerhizin, perillartine, naringin dihydrochalcone, neohesperidine dihydrochalcone, mogroside V, rubusoside, rubus extract, and rebaudioside A.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. Ser. No. 12/593,114, field onSep. 25, 2009, which is a national stage application of InternationalApplication No. PCT/CH2008/000135, filed 27 Mar. 2008, which claims thebenefit of the filing date under 35 U.S.C. 119(e) of U.S. ProvisionalPatent Application Ser. No. 60/909,143, filed 30 Mar. 2007 and U.S.Provisional Patent Application Ser. No. 60/962,515, filed 30 Jul. 2007,from which applications priority is claimed, and which applications arehereby incorporated herein by reference in their entireties.

TECHNICAL FIELD

Disclosed are compounds that allow to mask or block undesirableoff-notes in consumables and the method of blocking off-notes employingsaid compounds in consumables.

SUMMARY

Provided are the following:

(1) An off-note blocking compound selected from one or more of4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoic acid,4-(2,2,3-trimethylcyclopentyl)butanoic acid,4-(2,2,6-trimethylcyclohexyl)butanoic acid,3-(2,2,6-trimethylcyclohexyl)propanoic acid,2-(3,4-dimethylcyclohexyl)acetic acid,2-methyl-4-(2,6,6-trimethylcyclohex-2-enyl)butanoic acid,1-methyl-2-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropanecarboxylicacid; (E)-3-(1-(2,2,3-trimethylcyclopent-3-enyl)cyclopropyl)acrylicacid, (E)-4-(2,2,3-trimethylcyclohex-3-enyl)but-2-enoic acid,(E)-4-(2,2,3-trimethylcyclopentyl)but-3-enoic acid,(E)-4-(2,2,3-trimethylcyclopentyl)but-2-enoic acid,1-methyl-2-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropanecarboxylicacid, 2,2-dimethyl-4-(2,2,3-trimethylcyclopentyl)butanoic acid,2-((1,4-dimethylbicyclo[3.1.0]hexan-2-yl)methyl)-1-methylcyclopropanecarboxylicacid,2-(1-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropyl)propanoicacid, 2-(1-((1-methylbicyclo[3.1.0]hexan-2-yl)methyl)cyclopropyl)aceticacid, 3,3-dimethyl-4-(2,2,3-trimethylcyclopentyl)butanoic acid, and4-(1-methylbicyclo[3.1.0]hexan-2-yl)butanoic acid,(E)-1-(2-(1-methylbicyclo[3.1.0]hexan-2-yl)vinyl)cyclopropanecarboxylicacid,(E)-1-(2-(7-methylspiro[2.4]heptan-4-yl)vinyl)cyclopropanecarboxylicacid, (E)-4-(1-methylbicyclo[3.1.0]hexan-2-yl)but-3-enoic acid,(E)-4-(7-methylspiro[2.4]hept-6-en-4-yl)but-2-enoic acid,(E)-4-(7-methylspiro[2.4]heptan-4-yl)but-3-enoic acid,(E)-4-(7-methylspiro[2.4]heptan-4-yl)but-2-enoic acid,(E)-4-(8-methylspiro[2.5]oct-7-en-4-yl)but-2-enoic acid,1-(2-(7-methylspiro[2.4]heptan-4-yl)ethyl)cyclopropanecarboxylic acid,2-((6,7-dimethylspiro[2.4]heptan-4-yl)methyl)-1-methylcyclopropanecarboxylicacid,2-(1-((1,4-dimethylbicyclo[3.1.0]hexan-2-yl)methyl)cyclopropyl)propanoicacid, 2-(1-((1-methylbicyclo[3.1.0]hexan-2-yl)methyl)cyclopropyl)aceticacid, 2-(1-((7-methylspiro[2.4]heptan-4-yl)methyl)cyclopropyl)aceticacid, and 4-(7-methylspiro[2.4]heptan-4-yl)butanoic acid.

(2) The off-note blocking compound as described herein, including under(1), selected from one or more of 4-(2,2,3-trimethylcyclopentyl)butanoicacid, 4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoic acid,4-(2,2,6-trimethylcyclohexyl)butanoic acid,3-(2,2,6-trimethylcyclohexyl)propanoic acid4-propylcyclohexanecarboxylicacid, and 2-(3,4-dimethylcyclohexyl)acetic acid.

(3) A flavor composition comprising an off-note providing consumableingredient and one or more of the off-note blocking compounds listedherein, including under (1) and (2).

(4) A consumable comprising

-   -   a) one or more ingredient in a concentration sufficient to        provide an off-note, and    -   b) one or more of the off-note blocking compounds listed herein,        including under (1) and (2).

(5) A consumable as herein described, including under (4), wherein theone or more off-note providing ingredient is selected from one or moreof sweetener, artificial sweetener, beverage, chewing gum,nutraceutical, and pharmaceutical.

(6) A consumable of as herein described, including under (4), whereinthe one or more off-note providing ingredient is an artificial sweetenerselected from one or more of aspartame, acesulfame K, saccharin,sucralose, and sodium cyclamate.

(7) A consumable as herein described, including under (4), wherein theone or more off-note providing ingredient comprises a sweetener selectedfrom one or more of stevioside, swingle extract, glyccerhizin,perillartine, naringin dihydrochalcone, neohesperidine dihydrochalcone,mogroside V, rubusoside, rubus extract, and rebaudioside A

(8) A consumable as herein described, including under (4), wherein theoff-note providing ingredient comprises a consumable selected from oneor more of cocoa, coffee, caffeine, theobromine, diketopiperazines,vitamins, amino acids, vitamin B, casein, soy protein, ibuprofen,salicylic acid, glucoronolactone, acetaminophen, dextromethorphan,naringin, taurin, macrolide (including bioxin and erythomycin),paracetamol, acetylsalicylic acid, cimetidine, ranitidine, amoxicillin,acetominophen, cephalosporines, quassia, propylene glycol, triacetin,salts of potassium, salts of zinc, loperamide, limonin, flavonoides,isoflavones (including genistein and diadzein), polyphenol (includingcatechin and epicatechin), mint oil, D-menthol, hydrolysed vegetableprotein, bitter peptides, preservatives (including benzoic acid,potassium sorbate, polysorbate 80, sodium and potassium lactate, sodiumbenzoate), citric acid, quinine, urea (contained in chewing gums),essential oils (including thyme, sage, basil, mint), Maillard reactionproducts (including cyclic amines made from pyrrolidine/glucose,alanine/xylose, proline/sucrose or alanine/xylose, for examplediketopiperazines), beer, hops, humulone, trans-isohumulone, lupulone,and hulupone.

(10) A method of blocking off-notes in a consumable comprising admixingwith the consumable (a) one or more off-note providing ingredient in aconcentration sufficient to provide an off-note, and (b) one or morecompounds of the off-note blocking compounds listed herein, includingunder (1) and (2).

(11) A method as herein described, including under (10), wherein the oneor more off-note providing ingredient is selected from the groupconsisting of sweetener, artificial sweetener, beverage, chewing gum,nutraceutical, and pharmaceutical.

(12) A method as herein described, including under (10) and (11),wherein the off-note providing ingredient comprises one or moreartificial sweetener selected from aspartame, acesulfame K, saccharin,sucralose, and sodium cyclamate.

(13) A method of as herein described, including under (10) to (12),wherein the off-note providing ingredient comprises one or moresweetener selected from stevioside, swingle extract, glyccerhizin,perillartine, naringin dihydrochalcone, neohesperidine dihydrochalcone,mogroside V, rubusoside, rubus extract, and rebaudioside A.

(14) A method as herein described, including under (10) to (13), whereinthe off-note providing ingredient comprises one or more consumableselected from cocoa, coffee, caffeine, theobromine, diketopiperazines,vitamins, amino acids, vitamin B, casein, soy protein, ibuprofen,salicylic acid, glucoronolactone, acetaminophen, dextromethorphan,naringin, taurin, macrolide (including bioxin and erythomycin),paracetamol, acetolsalicilic acid, cimetidine, ranitidine, amoxicillin,acetominophen, cephalosporines, quassia, propylene glycol, triacetin,salts of potassium, salts of zinc, loperamide, limonin, flavonoides,isoflavones (including genistein and diadzein), polyphenol (includingcatechin and epicatechin), mint oil, D-menthol, hydrolysed vegetableprotein, bitter peptides, preservatives (including benzoic acid,potassium sorbate, polysorbate 80, sodium and potassium lactate, sodiumbenzoate), citric acid, quinine, urea (contained in chewing gums),essential oils (including thyme, sage, basil, mint), Maillard reactionproducts (including cyclic amines made from pyrrolidine/glucose,alanine/xylose, proline/sucrose or alanine/xylose, for examplediketopiperazines), beer, hops, humulone, trans-isohumulone, lupulone,and hulupone.

DETAILED DESCRIPTION

The off-note blocking compounds provided herein include the followingoff-note blocking compounds:4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoic acid,4-(2,2,3-trimethylcyclopentyl)butanoic acid,4-(2,2,6-trimethylcyclohexyl)butanoic acid,3-(2,2,6-trimethylcyclohexyl)propanoic acid,2-(3,4-dimethylcyclohexyl)acetic acid,2-methyl-4-(2,6,6-trimethylcyclohex-2-enyl)butanoic acid, and1-methyl-2-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropanecarboxylicacid. The chemical structures of these compounds are indicated below:

Without wishing to be bound by theory, the presence of 1 to 3cyclopropyl groups at the 5-ring or 6-ring part of the off-note blockingcompound and/or the linker linking the ring to the acid group tends toincrease the off-note blocking activity. According to certainembodiments, these compounds are particularly useful for blocking thebitter off-note blocking activity. Furthermore, 5-ring structures(cyclopentane) tend to have higher activities than the related 6-ringstructures (cyclohexane).

According to certain embodiments, the following groups of compounds mayalso be useful for off-note blocking in consumables:

One or more compound selected from(E)-3-(1-(2,2,3-trimethylcyclopent-3-enyl)cyclopropyl)acrylic acid,(E)-4-(2,2,3-trimethylcyclohex-3-enyl)but-2-enoic acid,(E)-4-(2,2,3-trimethylcyclopentyl)but-3-enoic acid,(E)-4-(2,2,3-trimethylcyclopentyl)but-2-enoic acid,1-methyl-2-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropanecarboxylicacid, 2,2-dimethyl-4-(2,2,3-trimethylcyclopentyl)butanoic acid,2-((1,4-dimethylbicyclo[3.1.0]hexan-2-yl)methyl)-1-methylcyclopropanecarboxylicacid,2-(1-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropyl)propanoicacid, 2-(1-((1-methylbicyclo[3.1.0]hexan-2-yl)methyl)cyclopropyl)aceticacid, 3,3-dimethyl-4-(2,2,3-trimethylcyclopentyl)butanoic acid, and4-(1-methylbicyclo[3.1.0]hexan-2-yl)butanoic acid.

The chemical structures of these compounds are shown below.

Structure Name

(E)-4-(2,2,3-trimethylcyclopentyl) but-3-enoic acid

(E)-4-(2,2,3-trimethylcyclopentyl) but-2-enoic acid

(E)-4-(2,2,3-trimethylcyclohex-3- enyl)but-2-enoic acid

2,2-dimethyl-4-(2,2,3- trimethylcyclopentyl)butanoic acid

3,3-dimethyl-4-(2,2,3- trimethylcyclopentyl)butanoic acid

4-(1-methylbicyclo[3.1.0]hexan- 2-yl)butanoic acid

1-methyl-2-((1,2,2- trimethylbicyclo[3.1.0]hexan-3- yl)methyl)cyclopropanecarboxylic acid

2-((1,4-dimethylbicyclo[3.1.0] hexan-2-yl)methyl)-1-methylcyclopropanecarboxylic acid

2-(1-((1-methylbicyclo[3.1.0] hexan-2-yl)methyl) cyclopropyl)acetic acid

2-(1-((1,2,2-trimethylbicyclo [3.1.0]hexan-3- yl)methyl)cyclopropyl)propanoic acid

(E)-3-(1-(2,2,3- trimethylcyclopent-3- enyl)cyclopropyl)acrylic acid

According to further embodiments, the following group of compounds mayalso be useful in off-note blocking in consumables:

(E)-1-(2-(1-methylbicyclo[3.1.0]hexan-2-yl)vinyl)cyclopropanecarboxylicacid,(E)-1-(2-(7-methylspiro[2.4]heptan-4-yl)vinyl)cyclopropanecarboxylicacid, (E)-4-(1-methylbicyclo[3.1.0]hexan-2-yl)but-3-enoic acid,(E)-4-(7-methylspiro[2.4]hept-6-en-4-yl)but-2-enoic acid,(E)-4-(7-methylspiro[2.4]heptan-4-yl)but-3-enoic acid,(E)-4-(7-methylspiro[2.4]heptan-4-yl)but-2-enoic acid,(E)-4-(8-methylspiro[2.5]oct-7-en-4-yl)but-2-enoic acid,1-(2-(7-methylspiro[2.4]heptan-4-yl)ethyl)cyclopropanecarboxylic acid,2-((6,7-dimethylspiro[2.4]heptan-4-yl)methyl)-1-methylcyclopropanecarboxylicacid,2-(1-((1,4-dimethylbicyclo[3.1.0]hexan-2-yl)methyl)cyclopropyl)propanoicacid, 2-(1-((1-methylbicyclo[3.1.0]hexan-2-yl)methyl)cyclopropyl)aceticacid, 2-(1-((7-methylspiro[2.4]heptan-4-yl)methyl)cyclopropyl)aceticacid, and 4-(7-methylspiro[2.4]heptan-4-yl)butanoic acid.

The chemical structures of these compounds are shown below.

Structure name

(E)-4-(7-methylspiro[2.4]hept- 6-en-4-yl)but-2-enoic acid

(E)-4-(8-methylspiro[2.5]oct- 7-en-4-yl)but-2-enoic acid

4-(7-methylspiro[2.4]heptan- 4-yl)butanoic acid

2-(1-((7-methylspiro[2.4]heptan- 4-yl)methyl)cyclopropyl)acetic acid

2-(1-((1-methylbicyclo[3.1.0] hexan-2-yl)methyl)cyclopropyl) acetic acid

2-((6,7-dimethylspiro[2.4] heptan-4-yl)methyl)-1-methylcyclopropanecarboxylic acid

1-(2-(7-methylspiro[2.4] heptan-4-yl)ethyl) cyclopropanecarboxylic acid

(E)-1-(2-(7-methylspiro[2.4] heptan-4-yl)vinyl) cyclopropanecarboxylicacid

2-(1-((1,4-dimethylbicyclo [3.1.0]hexan-2-yl)methyl)cyclopropyl)propanoic acid

(E)-4-(7-methylspiro[2.4] heptan-4-yl)but-3-enoic acid

(E)-4-(7-methylspiro[2.4] heptan-4-yl)but-2-enoic acid

(E)-4-(1-methylbicyclo[3.1.0] hexan-2-yl)but-3-enoic acid

(E)-1-(2-(1-methylbicyclo [3.1.0]hexan-2-yl)vinyl)cyclopropanecarboxylic acid

Most compounds can be synthesized easily by methods well known in theart, or as indicated below or in the examples herein.

Synthesis of 2-methyl-4-(2,6,6-trimethylcyclohex-2-enyl)butanoic acid:2-methyl-4-(2,6,6-trimethylcyclohex-2-enyl)butanoic acid can besynthesized in one step from2-methyl-4-(2,6,6-trimethylcyclohex-2-enyl)butanal analoguously to thesynthesis pathway as described by Ochiai et al. 1989, Journal of OrganicChemistry, 54(20), 4832-40.

Synthesis of 2-(3,4-dimethylcyclohexyl)acetic acid:2-(3,4-dimethylcyclohexyl)acetic acid can be synthesized from2-(3,4-dimethylphenyl)acetic acid as indicated below using hydrogen andplatinumoxide (H₂/PtO₂) as catalyst analoguously to the synthesispathway as described by Gault et al. (1958), Compt. rend., 246, 123-5.

Synthesis of1-methyl-2-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropanecarboxylicacid:

The compound can be synthesized starting from(1-methyl-2-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropyl)methanol(aka Javanol®) using Ruthenium(III) chloride and sodium periodateanaloguously to the synthesis pathway as described by Hart et al. 2003,Journal of Organic Chemistry 68(1), 187-190. Javanol® is commerciallyavailable from Givaudan, Vernier, Switzerland.

The synthesis of further compounds is described in examples 2a to 2dherein below. The remainder of compounds can be synthesized analogously,as will be apparent to the skilled person.

A receptor screen with concentration-response analysis was performed andfrom results inhibitory concentration (IC) IC₅₀ values can be calculatedby nonlinear regression using the function f(x)=(a-d)/(1+(x/C)^(nh))+d;with a=minimum signal, d=maximum signal, nh=hill coefficient, C=IC₅₀.and x=concentration of antagonist. IC₅₀ is the molar concentration of anantagonist which produces 50% of the maximum possible inhibitoryresponse for that antagonist. A more potent antagonist will have a lowerIC₅₀ value.

Most of the off-note blocking compounds disclosed herein, and inparticular, for example, 4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoicacid, 4-(2,2,3-trimethylcyclopentyl)butanoic acid,4-(2,2,6-trimethylcyclohexyl)butanoic acid,3-(2,2,6-trimethylcyclohexyl)propanoic acid,2-(3,4-dimethylcyclohexyl)acetic acid,2-methyl-4-(2,6,6-trimethylcyclohex-2-enyl)butanoic acid, and1-methyl-2-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropanecarboxylicacid have an IC₅₀ in the range of about 0.1 to 20 micromolar when testedwith the TAS2R44 bitter taste receptor.

For most food applications, a low IC₅₀ [micro molar] of 0.05 to 10 isdesirable, however, IC₅₀ of 10 to 25 are still good and above 25 mayalso still acceptable depending on the application.

Various food ingredients (including ingredients naturally contained infood or additives admixed to food including flavor ingredients) provideundesirable off-notes. Particularly undesirable off-notes are the bitteroff-notes, metallic off-notes, lingering, licorice-type and astringentoff-notes. The term off-note refers to an unpleasant after taste thatdevelops over time after consumption of consumables.

Other particular examples are the bitter and/or metallic and/orastringent and/or “artificial” off-notes and/or a cloyingly sweetoff-note (as opposed to the “cleaner” taste of sugar) that areassociated with a number of artificial sweeteners including aspartame,

Ace K, saccharin, sucralose, and sodium cyclamate. Sometimes theseoff-notes of artificial sweeteners are described collectively as bitteroff-notes.

Further examples of off-note providing ingredients are naturallyoccurring sweeteners including stevioside, swingle extract,glyccerhizin, perillartine, naringin dihydrochalcone, neohesperidinedihydrochalcone, mogroside V, rubusoside, rubus extract, andrebaudioside A

Still further examples of off-note providing ingredients include cocoa,coffee, caffeine, theobromine, diketopiperazines, vitamins, amino acids,vitamin B, casein, soy protein, ibuprofen, salicylic acid,glucoronolactone, acetaminophen, dextromethorphan, naringin, taurin,macrolide (including bioxin and erythomycin), paracetamol,acetolsalicilic acid, cimetidine, ranitidine, amoxicillin,acetominophen, cephalosporines, quassia, propylene glycol, triacetin,potassium, zinc, loperamide, limonin, flavonoides, isoflavones(including genistein and diadzein), polyphenol (including catechin andepicatechin), mint oil, D-menthol, hydrolysed vegetable protein, bitterpeptides, preservatives (including benzoic acid, potassium sorbate,polysorbate 80, sodium and potassium lactate, sodium benzoate), citricacid, quinine, urea (contained in chewing gums), essential oils(including thyme, sage, basil, mint), Maillard reaction products(including cyclic amines made from pyrrolidine/glucose, alanine/xylose,proline/sucrose or alanine/xylose, for example diketopiperazines), beer,hops, humulone, trans-isohumulone, lupulone, hulupone.

The addition of off-note blockers will block or mask the off-notes andmake them less apparent or unnoticeable. Artificial sweeteners willthereby loose their bitter/metallic taste and/or their cloyingly sweetlingering sweetness and instead taste more like actual sugar (sucrose).

Aspartame is the name for aspartyl-phenylalanine-1-methyl ester, adipeptide. It is known under various trademark names including Equal®,and Canderel®. In the European Union, it is also known under the Enumber (additive code) E951.

Acesulfame potassium (AceK) is the potassium salt of6-methyl-1,2,3-oxathiazine-4(3H)-one 2,2-dioxide, an N-sulfonylamide. Itis also known as Acesulfame K or AceK, or under various trademark namesincluding Sunett® and Sweet One®. In the European Union it is also knownunder the E number (additive code) E950.

Saccharin is the Na salt of 1,2-Benzisothiazol-3(2H)-one, 1,1-dioxide,an N-sulfonamide. It is also known under various trademark namesincluding Sweet'n low®.

Sucralose is the name for 6-dichloro-1,6-dideoxy-β-D-fructo-furanosyl4-chloro-4-deoxy-α-D-galactopyranoside, which is a chlorodeoxysugar. Itis also known by the trade name Splenda®. In the European Union, it isalso known under the E number (additive code) E955. Sucralose has anoff-note (also designated “aftertaste”) that is a lingeringliquorice-like off-note sometimes also described as bitter.

The off-note blockers can be added to consumables to block theundesirable off-notes of ingredients present in said consumables oradded to such consumables.

Flavor compositions for addition to consumables can be formed thatprovide the off-note blockers and an off-note providing ingredient foraddition to consumables, and optionally food grade excipients.Alternatively, the off note blockers can be directly added toconsumables.

In particular, the off-note blockers can be added to flavor compositionsor directly to consumables to block the undesirable off-notes ofoff-note providing ingredients including natural and artificialsweeteners added to such consumables.

Consumables include all food products, food additives, nutraceuticals,pharmaceuticals and any product placed in the mouth including chewinggum, oral care products, and oral hygiene products including but notlimited to, cereal products, rice products, tapioca products, sagoproducts, baker's products, biscuit products, pastry products, breadproducts, confectionery products, desert products, gums, chewing gums,mouthwash, dental floss, flavored or flavor-coated straws, flavor orflavor-coated food/beverage containers, chocolates, ices, honeyproducts, treacle products, yeast products, baking-powder, salt andspice products, savory products, mustard products, vinegar products,sauces (condiments), tobacco products, cigars, cigarettes, processedfoods, cooked fruits and vegetable products, meat and meat products,jellies, jams, fruit sauces, egg products, milk and dairy products,yoghurts, cheese products, butter and butter substitute products, milksubstitute products, soy products, edible oils and fat products,medicaments, beverages, carbonated beverages, alcoholic drinks, beers,soft drinks, mineral and aerated waters and other non-alcoholic drinks,fruit drinks, fruit juices, coffee, artificial coffee, tea, cocoa,including forms requiring reconstitution, food extracts, plant extracts,meat extracts, condiments, nutraceuticals, gelatins, pharmaceutical andnon-pharmaceutical gums, tablets, lozenges, drops, emulsions, elixirs,syrups and other preparations for making beverages, and combinationsthereof.

For example, in consumables containing potassium, the off-note blockermay be added to suppress the bitterness and the metallic off-noteassociated with potassium.

In coffee and cocoa products, the off-note blocker may be added tosuppress the bitterness associated with caffeine, theobromine, and/ordiketopiperazines present in said products.

In cheese products, in particular in enzyme-modified cheese products,the off-note blocker may be added to suppress the bitterness associatedwith bitter peptides present in said cheese products.

In soy products, the off-note blocker may be added to suppress thebitterness and beany off-notes associated with peptides, isoflavonessuch as genistein and diadzein present in said products.

In HVP (hydrolysed vegetable protein) products, the off-note blocker maybe added to suppress the bitterness associated with bitter peptidespresent in said products.

In functional ingredients used in fortified foods, the off-note blockermay be added to suppress the bitterness associated with vitamins andamino acids present in said products.

In pharmaceuticals, the off-note blocker may be added to suppress thebitterness associated with actives or bitter additives present in saidproducts.

In consumables containing solvents, the off-note blocker may be added tosuppress the bitterness associated with propylene glycol, triacetin, orethanol present in said products.

In citrus products, the off-note blocker may be added to suppress thebitterness associated with naringin present in said products.

In neutraceuticals and herb medicines, the off-note blocker may be addedto suppress the bitterness associated with actives or additives presentin said products.

In consumables containing polyphenols such as catechin and epicatechin,the off-note blocker may be added to suppress the bitterness associatedwith these ingredients.

In consumables containing preservatives such as potassium sorbate,polysorbate 80, sodium and potassium lactate, sodium benzoate, theoff-note blocker may be added to suppress the bitterness associated withsaid preservatives.

In consumables containing zinc and other mineral supplements, theoff-note blocker may be added to suppress the bitterness and metallicoff-notes associated with said these mineral supplements.

In consumables containing mint oil or menthol (e.g. D-menthol) andcitric acid of above 7%, the off-note blocker may be added to suppressthe bitterness associated with this combination of ingredients.

In consumables containing quinine, the off-note blocker may be added tosuppress the bitterness associated with quinine.

In consumables containing artificial sweeteners (e.g. aspartame,saccharin, acesulfameK, sucralose, cyclamate), for example beverages, aoff-note blocker may be added to suppress the bitterness associated withartificial sweeteners.

In chewing gums, particular dental-type chewing gums, the off-noteblocker may be added to suppress the bitterness associated with ureacontained in chewing gums.

In consumables containing essential oils (e.g. thyme, sage, basil,mint), the off-note blocker may be added to suppress the bitternessassociated with these essential oils.

In consumables containing vegetables or herbs or their extracts, theoff-note blocker may be added to suppress the bitterness associated withthese ingredients.

In consumables containing Maillard reaction products (i.e. cyclic aminesmade from proline/sucrose or alanine/xylose, e.g. diketopiperazines),the off-note blocker may be added to suppress the bitterness associatedwith Maillard reaction products.

In beer and consumables containing beer or hops, the off-note blockermay be added to suppress the bitterness associated with hops.

EXAMPLES

The following examples are set forth to describe the off-note blockingcompounds in further detail and to illustrate the methods of employingthe off-note blocking compounds to block or otherwise mask off-notes inconsumables. The examples are illustrative and should not be construedas limiting the compounds, consumables or methods in any manner.

Example 1 Sensorial Evaluation in Various Consumables

Off-note blockers as herein described are tested by panels of 6 to 10bitter sensitive panelists.

Panelists are asked to describe the differences in off-notes and bitternotes between the product with 0.001% (wt/wt) off-note blocker unlessotherwise stated and a control without off-note blocker.

A) Aspartame/Acesulfame-K Containing Diet Energy Drink

The diet energy drink contained taurin, acesulfame K, aspartame,sucralose, glucuronolacton, caffeine, B-group vitamins (Niacin,pantothenic acid, B6, B12), aroma, sucrose, glucose, colours

The sample containing the off-note blocker is found to be less bittercompared to the control.

B) Sucrose/Glucose-Sweetened Energy Drink

The diet energy drink contains taurin, glucuronolacton, caffeine,B-group vitamins (Niacin, pantothenic acid, B6, B12), aroma, sucrose,glucose, colours.

The sample containing the off-note blocker is found to have lessoff-notes, to be less bitter, and less astringent compared to thecontrol.

C) Iced low-sugar coffee

The sample containing the off-note blocker is found to be less bitter,and less astringent compared to the control.

D) Commercial Vanilla Flavored Nutritional Drink

Vanilla flavored nutritional drink containing calcium caseinate, soyprotein isolate, sodium caseinate, vitamins and minerals.

The sample containing the off-note blocker is found to be less chalky,to have reduced protein/vitamin induced off-notes notes, and to be lessastringent compared to control.

E) Saccharin Sweetened Cola Soft Drink

The sample containing the off-note blocker is found to be less bitterand to have a reduced after taste compared to the control.

F) Loperamide Containing Mint-Flavored Pharmaceutical Syrup

The syrup contained 1 mg loperamide HCl per 7.5 ml serving. The off-noteblocker is used in a concentration of 0.004% (wt/wt).

The sample containing the off-note blocker is found to be less bitterwith especially the lingering bitter after taste reduced.

G) Daytime Cough Syrup

The daytime cough syrup contains 325 mg acetaminophen, 10 mgdextromethorphan HBr, 5 mg phenylephrine HCl per 15 ml serving.

The sample containing the off-note blocker is found to be less bitter.

H) Dark Chocolate

The sample containing the off-note blocker is found to be less bitter.

I ) Baking Chocolate (100% Cocoa, Unsweetened)

The off-note blocker is used in a concentration of 0.002% (wt/wt).

The sample containing the off-note blocker is found to be less metallic,less bitter, especially the alkaloid/caffeine-like bitterness is reducedwhile the upfront, warm, woody bitterness was retained.

J) Overcooked Coffee

Coffee is brewed and cooked on a burner for 3 hours. The off-noteblocker is used in a concentration of 0.0005% (wt/wt).

The sample containing the off-note blocker is found to be less bitter.

K) Aspartame/Acesulfame-K Sweetened Plain Nonfat Yoghurt

The yoghurt contained 0.0193% (wt/wt) and acesulfame-K 0.0083% (wt/wt).The off-note blocker is used in a concentration of 0.00175% (wt/wt).

The sample containing the off-note blocker is found to have lessoff-notes compared to control.

L) Aspartame/Acesulfame-K Sweetened Cola Soft Drink

The off-note blocker is used in a concentration of 0.0063% (wt/wt).

Cola soft drink % (by weight) Sodium Benzoate 0.026 Aspartame 0.043Acesulfame-K 0.017 Caffeine 0.011 Phosphoric Acid (85%) 0.043 CitricAcid (50% cut in water) 0.017 Caramel Color 0.085 Water Balance to 100

The sample containing the off-note blocker is found to have lessoff-notes than the control.

M) Sucralose Sweetened Cola Soft Drink

The off-note blocker is used in a concentration of 0.0035% (wt/wt).

sucralose cola soft drink % (by weight) Sodium Benzoate 0.03 Sucralose(25% cut in water) 0.06 Caffeine 0.01 Phosphoric Acid (85%) 0.08 CaramelColor 0.09 Water Balance to 100

The sample containing the off-note blocker is found to have lessoff-notes and be less bitter than the control.

Examples 2a-2xy Synthesis of Off-Note Blocking Compounds Example 2aSynthesis of 4-(2,2,3-trimethylcyclopentyl)butanoic acid:

4-(2,2,3-trimethylcyclopentyl)butanoic acid was synthesized in threesteps; the first step reacts2-(2,2,3-trimethylcyclopent-3-en-1-yl)acetaldehyde (aka campholenicaldehyde) to ethyl 4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoate, inthe second step the latter was subjected to a hydrogenation procedure toform ethyl 4-(2,2,3-trimethylcyclopentyl)butanoic acid, and in the thirdstep, from the latter, 4-(2,2,3-trimethylcyclopentyl)butanoic acid wasformed in presence of sodium hydroxide (NaOH) and tetrahydrofuran (THF).

Step 1 (Formation of ethyl4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoate):

To a flame-dried 250 ml round bottom flask campholenic aldehyde (10.0 g,65.7 mmol) and methyl tertiary-butyl ether (MTBE) (100 ml) were added togive a solution and the flask was slowly cooled to 0° C. in an ice waterbath. (Carbethoxymethylene)triphenyl-phosphorane (22.8 g, 66.0 mmol) wasslowly added to the solution. The reaction mixture was stirred at 0° C.for 20 min then stirred at room temperature for about 24 h. After thereaction was completed, the reaction mixture was reduced to half itsvolume in vacuo and 100 ml of hexanes was added. The flask containingthe reaction mixture and hexanes was then placed in an ice bath for 30min. The reaction mixture was then filtered through a plug of filterpaper, celite, silica, and sand and washed three times, first withhexanes, then 9:1 and then 1:1 Hexanes/MTBE. The combined eluates wereconcentrated in vacuo and purifed via flash column chromatography (1:8Hexanes/Ethyl Acetate (AcOEt)) to give ethyl4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoate (13.1 g, 89%) as an oil.The NMR data is indicated below.

¹H NMR (300 MHz, CDCl3) δ 6.92 (dd, J=10.5, 7.5 Hz, 1H), 5.79 (d, J=15.6Hz, 1H), 5.13 (s, 1H), 4.13 (m, 2H), 2.3 (m, 2H), 2.1 (m, 1H), 1.8 (m,2H), 1.52 (s, 3H), 1.24 (t, J=15, 3H), 0.91 (s, 3H), 0.71 (s, 3H) ; ¹³CNMR (75 MHz, CDCl3) δ 166.1, 148.6, 147.8, 121.5, 121.3, 59.6, 48.9,46.2, 35.2, 32.9, 25.5, 19.5, 14.0, 12.2; MS m/z 222.

Step 2—Hydrogenation Procedure:

To a three necked round bottom flask with a stir bar flushed withnitrogen, Degussa type palladium on charcoal Pd/C (1.5 g, 10% Pd) wasadded as catalyst. The catalyst was covered with distilled water (8 ml)and ethyl 4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoate/ethyl4-(2,2,3-trimethylcyclopentenyl)butanoic acid (10.0 g, 44.9 mmol) inEthyl Acetate (AcOEt) (200 ml) was added to the reaction flask. Theflask was flushed with hydrogen and the reaction was run under hydrogenballoons that were refilled as needed. After 17 h at room temperaturethe reaction was found to be completed by gas chromatography and massspectrometry (GCMS). The reaction mixture was filtered through a celiteplug. The organic layers were concentrated in vacuo and purified viaflash column chromatography AcOEt/Hexanes (gradient 0 to 40%) to giveethyl 4-(2,2,3-trimethylcyclopentyl)butanoate/ethyl4-(2,2,3-trimethylcyclopentyl)butanoic acid as a colorless oil (7.48 g,74%). The NMR data is indicated below.

¹H NMR (300 MHz, CDCl3) δ 4.18 (q, J=7.14, 2H), 2.34 (m, 2H), 1.82 (m,3H), 1.56 (m, 5H), 1.30 (t, J=7.2 , 3H), 0. (m, 2H), 0.99 (s, 3H), 0.86(d, J =5.4, 2H), 0.52 (s, 3H); ¹³C NMR (75 MHz, CDCl3) δ 173.8, 60.1,50.6, 45.2, 42.2, 34.8, 30.1, 30.0, 28.1, 25.6, 24.3, 14.3, 14.2, 13.8;MS m/z 226.

Step 3—Saponification Reaction

Ethyl 4-(2,2,3-trimethylcyclopentyl)butanoate (5 g, 22 mmol) was addedto THF (25 ml) into a round bottom flask. Aqueous 1 N NaOH (25 ml) wasthen added to the flask and the reaction mixture was refluxed at 110° C.for 8 h. Upon completion of the reaction, the reaction mixture wasdiluted with 1 N NaOH (25 ml) and the aqueous layer washed twice withMTBE (50 ml×2). The aqueous layer was treated with aqueous 1.0 N HCluntil it reached a pH of about 3 (for example from 2 to 4), thenextracted three times with AcOEt (50 ml×3). The combined AcOEt extractswere concentrated in vacuo and purified via flash column chromatographywith (0-40% gradient) to give 4-(2,2,3-trimethylcyclopentyl)butanoicacid as a colorless oil (3.82 g, 87%). The NMR data is indicated below.

¹H NMR (300 MHz, CDCl3) δ 11.22 (br s, 1H), 2.37 (m, 2H), 1.75 (m, 3H),1.49 (m, 4H), 1.17 (m, 3H), 0.86 (s, 3H), 0.84 (d, J=6.9 Hz, 3H), 0.52(s, 3H); 13C NMR (75 MHz, CDCl3) δ 179.9, 50.6, 45.2, 42.2, 34.4, 30.1,30.0, 28.1, 25.0, 24.0, 14.3, 13.8; MS m/z 198.

Example 2b Synthesis of 4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoicacid:

4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoic acid was synthesized intwo steps; the first step reacts campholenic aldehyde to ethyl4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoate as describedherein-above, and in the second step, from the latter,4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoic acid was formed inpresence of NaOH and THF (see reaction below).

Ethyl 4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoate (2.0 g, 9.0 mmol)was added to THF (10 ml) in a 50 ml round bottom flask. Aqueous 1 N NaOH(10 ml) was then added to the flask and the reaction mixture wasrefluxed at 83° C. for 19 h. Upon completion of the reaction, thereaction mixture was diluted with 1 N NaOH (10 ml) and the aqueous layerwas washed twice with MTBE (10 ml×2). The aqueous layer was treated withaqueous 1.0 N HCl until it reached a pH of about 3 (for example from 2to 4), then extracted three times with AcOEt (10 ml×3). The combinedAcOEt extracts were concentrated in vacuo and purified via flash columnchromatography with MTBE/Hexanes (5-20% gradient) to give4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoic acid (1.58 g) as aviscous oil (1.58 g, 90%). The NMR data is indicated below.

¹H NMR (300 MHz, CDCl3) δ 11.47 (br. s, 1H), 7.15 (m, 1H), 5.90 (d,J=2.8 Hz 1H), 5.23 (s, 1H), 2.39 (m, 3H), 1.91 (m, 2H), 1.62 (s, 3H),1.20 (s, 3H), 0.827 (s, 3H); ¹³C NMR (75 MHz, CDCl3) δ 171.3, 152.2,148.3, 121.9, 120.9, 49.1, 46.9, 35.4, 33.4, 25.8, 19.7, 12.5; MS m/z194.

Example 2c Synthesis of 4-(2,2,6-trimethylcyclohexyl)butanoic acid:

4-(2,2,6-trimethylcyclohexyl)butanoic acid was synthesized in threesteps starting from 2-(2,6,6-trimethylcyclohex-1-enyl)acetaldehyde,analogous to the 3-step procedure described for4-(2,2,3-trimethylcyclopentyl)butanoic acid herein-above, with theexception that AcOEt was replaced by methanol (MeOH) in step 2, asindicated below. The first step forms (E)-ethyl4-(2,6,6-trimethylcyclohex-1-enyl)but-2-enoate, the second step formsethyl 4-(2,2,6-trimethylcyclohexyl)butanoate, the third forms4-(2,2,6-trimethylcyclohexyl)butanoic acid. The majority of the productcomprises (E)-4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoic acid), and(Z)-4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoic acid) is present in aconcentration of up to 10% (w/w) in the isomeric mixture.

Example 2d Synthesis of 3-(2,2,6-trimethylcyclohexyl)oropanoic acid

3-(2,2,6-trimethylcyclohexyl)propanoic acid was synthesized in threesteps starting from 2,6,6-trimethylcyclohex-1-enecarbaldehyde, analogousto the 3-step procedure described for4-(2,2,6-trimethylcyclohexyl)butanoic acid and4-(2,2,3-trimethylcyclopentyl)butanoic acid herein-above, as indicatedbelow.

The first step formed (E)-ethyl3-(2,6,6-trimethylcyclohex-1-enyl)acrylate, the second step formed ethyl3-(2,2,6-trimethylcyclohexyl)propanoate, the third formed the product3-(2,2,6-trimethylcyclohexyl)propanoic acid.

Example 3 TAS2R44 Bitter Taste Receptor Assay for IC 50 Determination 1)Generation of Human TAS2R44 Expression Vector

The full length gene of human TAS2R44 was amplified by polymerase chainreaction (PCR) using gene-specific primers that span the entire codingregion as described in WO 2004/029087.

The TAS2R44 cDNA was subcloned into an expression cassette based oneither of the following plasmids/expression vectors: pcDNA3.1Zeo(Invitrogen). These vectors contain within their multiple cloning sitesthe nucleotide sequence coding for the first 45 amino acids of the ratsomatostatin receptor subtype 3 (RSS tag) to facilitate cell surfacetargeting of the transgene (SEQ ID #4) and the nucleotide sequencecoding for the herpes simplex virus (HSV) glycoprotein D epitope (HSVepitope in aminoterminal to carboxyterminal direction, HSV tag). (SEQ ID#3) for facilitating immunocytochemical detection.

The TAS2R44 construct contains RSS tag, TAS2R44, and the HSV tag whichare fused in frame to allow translation into the receptor protein andthe resulting receptor cDNA.

This transfected expression vector is called pcDNA3.1Zeo-TAS2R44 (SEQ ID#1) and allows for expression of the TAS2R44 protein (SEQ ID #2).

2) Generation of a Cell Line Stably Expressing Gα16-Gustducin44 andTAS2R44

A cell line that stably expresses the human bitter taste receptor(TAS2R44) was generated by transfecting pcDNA3.1Zeo-TAS2R44 intoHEK293T/Gα16-gustducin 44 cells (both formed as described in under 1)above. The host cell line HEK-293T is commercially available from theAmerican Tissue Culture Collection (catalog #CRL-1573).

Transfection was performed as follows:

On day 0, the HEK293T Gα-16-gustducin44 cells were seeded in a 6-wellplate at a density of 900,000 cells per well and grown over night inselective growth medium (DMEM with 10% (v/v) heat-inactivated fetalbovine serum, 2 mM L-glutamine, 100 units/ml penicillin, 100 μg/mlstreptomycin, 200 μg/m1 G418 and 200 μg/ml zeocin). On day 1, the mediumwas exchanged with 2 ml of antibiotic-free and serum-free growth medium.10 μl Lipofectamine 2000 was dissolved in 250 μl DMEM and incubated for5 minutes at room temperature. In parallel, 4 μg TAS2R44 vector DNA weredissolved in 250 μl DMEM. These two resulting solutions are mixed andincubated for 20 minutes at room temperature before they are added tothe cells into the cell culture medium. After 4 hours, the medium isreplaced with antibiotic-free, serum-containing growth medium. The cellswere incubated in humidified atmosphere (37° C., 5% CO₂).

After 24 hours, the cells were re-plated in selective growth medium andwere further incubated in a humidified atmosphere (37° C., 5% CO₂).

After 2 to 4 weeks of culture (replacing medium as necessary),zeocin-resistant colonies were selected and expanded.

The selected clone was tested successfully for functional expression ofTAS2R44.

3) Fluo-4 Calcium Assay

Fluo-4 AM (Invitrogen) is a fluorescent indicator of intracellularcalcium dynamics (change in concentration) and allows monitoring changesin the calcium concentration, particularly an increase in response toreceptor activation occurring after agonist exposure.

At day 0, the HEK293T cell line stably expressing Gα16-gustducin44 andTAS2R44 formed as described under 2) was seeded in antibiotic-freegrowth medium (standard DMEM with 10% (v/v) heat-inactivated fetalbovine serum, 2 mM L-glutamine standard DMEM with 10% (v/v)heat-inactivated fetal bovine serum, 2 mM L-glutamine, 100 units/mlpenicillin, and 100 μg/ml streptomycin) into black wall/clear bottom96-well plates, coated with poly(ethylenimine) (0.005% v/v) at aconcentration of 15,000 cells per well and incubated for 48 hours inhumidified atmosphere (37° C., 5% CO₂).

At the time of the assay, the growth medium was discarded and the cellswere further incubated in a humidified atmosphere (37° C., 5% CO₂) for 1hour with 50 μl of loading buffer consisting of 1.5 μM Fluo-4 AM and 2.5μprobenicid (Sigma-Aldrich) in DMEM.

Afterwards, the 96-well plate was washed 5 times with 200 μl of assaybuffer (130 mM NaCl5 mM KCl 10 mM HEPES, 2 mM CaCl₂, and 5 mM dextrose,pH 7.4) per well, using an automated plate washer (BioTek). The platewas further incubated for 30 minutes at room temperature in the dark toallow for complete de-esterification of the Fluo-4. Afterwards the platewas washed 5 times with 200 μl of assay buffer per well, andreconstituted with 180 μl of assay buffer per well.

For assay reading, the plate was placed in a Fluorometric Imaging PlateReader (FLIPR) (FLIPR-TETRA™, Molecular Devices), and receptoractivation was initiated by addition of 20μl of a tenfold concentratedagonist stock solution (to give the desired agonist end concentrationwhen added to the 180 microliter assay buffer volume), which wasprepared in assay buffer.

Fluorescence was continuously monitored for 20 seconds to give a signalbaseline (averaged to give F₀) prior to agonist addition and for 120seconds after agonist addition. The change in signal divided by F₀ givesΔF/F₀ indicated in the table, with ΔF being the maximum signal occurringwithin the 120 seconds minus the minimum signal (occurring within the120 seconds after agonist addition.

All data was collected from at least two independent experiments eachcarried out in triplicate.

A concentration-response analysis was performed and IC₅₀ values werecalculated by nonlinear regression using the functionf(x)=(a-d)/(1+(x/C)^(nh))+d; with a=minimum signal, d=maximum signal,nh=hill coefficient, C=IC₅₀, and x=antagonist concentration. IC₅₀ is themolar concentration of an antagonist which produces 50% of the maximumpossible effective/inhibitory response for that antagonist. A morepotent antagonist will have a lower IC₅₀ value.

The obtained calcium signals were corrected for the response of cellsexpressing only the G Protein α subunit (Gα16-gustducin44) andnormalized to the fluorescence of cells prior to the stimulus usingΔF/F0 (Fmax-Fmin/F0).

Example 4 Determination of IC50 of Off-Note Blockers, Saccharin

The following off-note blockers were tested:4-(2,2,3-trimethylcyclopentyl)butanoic acid,4-(2,2,6-trimethylcyclohexyl)butanoic acid,3-(2,2,6-trimethylcyclohexyl)propanoic acid,2-(3,4-dimethylcyclohexyl)acetic acid,2-methyl-4-(2,6,6-trimethylcyclohex-2-enyl)butanoic acid, and1-methyl-2-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropanecarboxylicacid.

The method was performed as described in example 3, using saccharin asagonist.

The cells are exposed to a constant concentration of saccharin (0.5 mM)and to a set of different concentrations of the off-note blocker. Afluo-4 calcium assay was performed as described above in example 3 andgave an IC₅₀ [micro molar] within the range of 0.05 to 25.

This means that the off-note blockers inhibited the response of theTAS2R44 bitter taste receptor and will be useful to block bitter taste.

Example 5 Determination of IC50 for Off-Note Blockers, Acesulfame K

The method was performed as described in example 4, exchanging saccharinfor Acesulfame K (0.8 mM) as agonist. An IC₅₀ within the same range wasdetermined. This means that the off-note blockers inhibited the responseof the TAS2R44 bitter taste receptor and will be useful to block bittertaste.

Example 6 Determination of IC50 for Off-Note Blockers, Sucralose

The method was performed testing4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoic acid as described inexample 4, exchanging saccharin for sucralose (50 mM) as agonist. AnIC₅₀ within the same range was determined. This means that the off-noteblockers inhibited the response of the TAS2R44 bitter taste receptor andwill be useful to block bitter taste.

Example 7 Sensory Evaluation of4-(2,2,3-trimethylcyclopent-3-enyl)but-2-enoic acid,

The bitter blocker (13 ppm) was tested in water with sucralose inconcentrations of 1 mM, 3 mM, and 7 mM by a panel of bitter sensitiveindividuals (15) in two replications per each concentration. The sampleswere compared to negative controls without bitter blocker and panelistswere instructed to choose the less bitter sample in a forced choicetest.

The bitter blocker was found to significantly reduce bitterness (22/30panelists chose the sample with bitter blocker as less bitter).

The sequences employed in the constructs and methods described-hereincan be found in the sequence listing herein-below.

While the compounds, consumable and methods have been described above inconnection with illustrative embodiments, it is to be understood thatother similar embodiments may be used or modifications and additions maybe made to the described embodiments for performing the same functionwithout deviating therefrom. Further, all embodiments disclosed are notnecessarily in the alternative, as various embodiments of the inventionmay be combined to provide the desired characteristics. Variations canbe made by one having ordinary skill in the art without departing fromthe spirit and scope of the invention. Therefore, the compounds,consumables and methods should not be limited to any single embodiment,but rather construed in breadth and scope in accordance with therecitation of the attached claims.

1. An off-note blocking compound selected from one or more of4-(2,2,6-trimethylcyclohexyl)butanoic acid,3-(2,2,6-trimethylcyclohexyl)propanoic acid,2-(3,4-dimethylcyclohexyl)acetic acid,2-methyl-4-(2,6,6-trimethylcyclohex-2-enyl)butanoic acid,1-methyl-2-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropanecarboxylicacid;(E)-3-(1-(2,2,3-trimethylcyclopent-3-enyl)cyclopropyl)acrylic acid,(E)-4-(2,2,3-trimethylcyclohex-3-enyl)but-2-enoic acid,(E)-4-(2,2,3-trimethylcyclopentyl)but-3-enoic acid,(E)-4-(2,2,3-trimethylcyclopentyl)but-2-enoic acid,1-methyl-2-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropanecarboxylicacid, 2,2-dimethyl-4-(2,2,3-trimethylcyclopentyl)butanoic acid,2-((1,4-dimethylbicyclo[3.1.0]hexan-2-yl)methyl)-1-methylcyclopropanecarboxylicacid,2-(1-((1,2,2-trimethylbicyclo[3.1.0]hexan-3-yl)methyl)cyclopropyl)propanoicacid, 2-(1-((1-methylbicyclo[3.1.0]hexan-2-yl)methyl)cyclopropyl)aceticacid, 3,3-dimethyl-4-(2,2,3-trimethylcyclopentyl)butanoic acid, and4-(1-methylbicyclo[3.1.0]hexan-2-yl)butanoic acid,(E)-1-(2-(1-methylbicyclo[3.1.0]hexan-2-vinyl)cyclopropanecarboxylicacid, (E)-1-(2-(7-methylspiro[2.4]heptan-4-vinyl)cyclopropanecarboxylicacid, (E)-4-(1-methylbicyclo[3.1.0]hexan-2-yl)but-3-enoic acid,(E)-4-(7-methylspiro[2.4]hept-6-en-4-yl)but-2-enoic acid,(E)-4-(7-methylspiro[2.4]heptan-4-yl)but-3-enoic acid,(E)-4-(7-methylspiro[2.4]heptan-4-yl)but-2-enoic acid,(E)-4-(8-methylspiro[2.5]oct-7-en-4-yl)but-2-enoic acid,1-(2-(7-methylspiro[2.4]heptan-4-yl)ethyl)cyclopropanecarboxylic acid,2-((6,7-dimethylspiro[2.4]heptan-4-yl)methyl)-1-methylcyclopropanecarboxylicacid,2-(1-((1,4-dimethylbicyclo[3.1.0]hexan-2-yl)methyl)cyclopropyl)propanoicacid, 2-(1-((1-methylbicyclo[3.1.0]hexan-2-yl)methyl)cyclopropyl)aceticacid, 2-(1-((7-methylspiro[2.4]heptan-4-yl)methyl)cyclopropyl)aceticacid, and 4-(7-methylspiro[2.4]heptan-4-yl)butanoic acid.